Measurement method of denucleation amount

A measurement method and quantity measurement technology, applied in the direction of measuring devices, botany equipment and methods, biochemical equipment and methods, etc., can solve the problems of cell development damage, uncertain amount of enucleation, etc., so as to improve the efficiency of enucleation and solve the problem of Effects of cellular developmental damage

Active Publication Date: 2018-03-30
NANKAI UNIV
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Problems solved by technology

[0003] The technical problem mainly solved by the present invention is to provide a method for measuring the amount of enucleation, which can solve the problem of the uncertainty of the enucleation amount in the blind aspiration enucleation operation and cause great damage to cell development

Method used

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  • Measurement method of denucleation amount
  • Measurement method of denucleation amount

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Embodiment Construction

[0012] The cells used in this example are domestic pig oocytes taken from a local slaughter house. The method of obtaining oocytes is as follows:

[0013] After the ovaries were taken out from the slaughterhouse, they were transported to the laboratory in a thermos flask containing 35° to 37° normal saline within two hours. Then immediately wash twice with 37° sterile normal saline containing 100IU / L penicillin and 50mg / L streptomycin. Oocytes are extracted from small sacs with a diameter of 2 to 6 mm on the ovary. The extracted cells are washed three times with TL-Hepes-PVA and then cultured in vitro maturation (IVM) in an incubator at 39° and a carbon dioxide concentration of 5%. 42 hour. After IVM, the cells were degloved with 0.1% hyaluronidase. Finally, the cells were washed three times with M199 to obtain the used cells.

[0014] The experimental operation of the present invention is performed based on the self-developed micro-operating system NK-MR601, which connects the...

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Abstract

The invention discloses a measurement method of denucleation amount. The method is characterized in that cytoplasm in an entry needle can be automatically gathered to change the primary strip shape into an irregular ball shape after being released to an operation solution for a while during the blind absorbing denucleation process; the size of the cytoplasm ball can be measured to accurately obtain the denucleation amount. According to the method, the problem that the denucleation amount cannot be accurately estimated in the blind absorbing denucleation process is solved.

Description

Technical field [0001] The invention relates to the technical field of somatic cell nuclear transfer. In particular, it relates to a method for measuring the amount of denuclearization. Background technique [0002] Enucleation is to completely remove the main genetic material in the oocyte, so as to ensure that the individual developed from the transferred nucleus has the highest genetic characteristics similar to the nucleus animal (clone animal). The amount of denucleation directly affects the results of later cell development. If the enucleation is incomplete, it can lead to aneuploidy of the recombinant embryonic chromosomes, resulting in polyploidy, abnormal cleavage, developmental obstruction and early embryonic death. If the nucleation is too much, it will accelerate the abnormal differentiation of recombinant embryos and affect embryo development. Blind suction is a commonly used method of enucleation. The cytoplasm of the polar body and 1 / 4-1 / 3 of the cytoplasm below...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/873G01N33/483
CPCC12N15/873G01N33/4833
Inventor 赵新冯泽阳刘曜玮孙明竹崔茂盛
Owner NANKAI UNIV
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